Zhiyu Zhang et al.Quantitative proteomic analysis of glycosylated proteins enriched from urine samples with magnetic ConA nanoparticles identifies potential biomarkers for small cell lung,Journal of Pharmaceutical and Biomedical Analysis,Available online 31 August 2021, 114352

ABSTRACT

Lung cancer has high morbidity and mortality and small cell lung cancer (SCLC) is a highly invasive malignant tumor with a very unfavorable survival rate. Early diagnosis and treatment can result in better prognosis for the SCLC patients but current diagnostic methods are either invasive or incapable for large-scale screen. Therefore, discovering biomarkers for early diagnosis of SCLC is of importance. In this work, we covalently coupled Concanavalin A (ConA) to functionalized magnetic nanoparticles to obtain magnetic ConA-nanoparticles (ConA-NPs) for the enrichment of glycosylated proteins. We then purified glycosylated proteins in 36 urine samples from 9 healthy controls, 9 SCLC patients, 9 lung adenocarcinoma (LUAD) patients, and 9 squamous cell lung carcinoma (LUSC) patients. The purified glycosylated proteins were digested and analyzed by LC-MS/MS for identification and quantification. Among the 398 identified proteins, 20, 15, and 1 glycosylated protein(s), respectively, were upregulated in the urine of SCLC, LUAD, and LUSC patients. Immunoblotting experiments further demonstrated that cathepsin C and transferrin were significantly upregulated in the ConA-NP purified urine of SCLC patients. This work suggests that glycosylated cathepsin C and transferrin might be able to serve as potential biomarkers for the noninvasive diagnosis of SCLC patients.

摘要

肺癌具有较高的发病率和死亡率，小细胞肺癌（SCLC）是一种侵袭性很强的恶性肿瘤，生存率非常低。早期诊断和治疗可以改善小细胞肺癌患者的预后，但目前的诊断方法要么是侵入性的，要么不能进行大规模筛查。因此，发现SCLC早期诊断的生物标志物具有重要意义。在这项工作中，我们将刀豆球蛋白A（ConA）与功能化磁性纳米颗粒(PuriMag G-COOH)共价偶联，以获得用于富集糖基化蛋白质的磁性ConA纳米颗粒（ConA NPs）。然后我们从9名健康对照者、9名小细胞肺癌患者、9名肺腺癌（LUAD）患者和9名肺鳞状细胞癌（LUSC）患者的36份尿液样本中纯化糖基化蛋白。纯化的糖基化蛋白经LC-MS/MS消化和分析，用于鉴定和定量。在398种已鉴定的蛋白中，SCLC、LUAD和LUSC患者的尿液中分别有20、15和1种糖基化蛋白上调。免疫印迹实验进一步证明，小细胞肺癌患者的ConA NP纯化尿液中组织蛋白酶C和转铁蛋白显著上调。这项研究表明，糖基化组织蛋白酶C和转铁蛋白可以作为小细胞肺癌患者无创性诊断的潜在生物标志物。

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